Synthetic polymers scaffolds often need to be coated with extracellular matrix (ECM) proteins to improve cell adhesion. For cultivated meat applications, coating should be avoided since it is necessary to eliminate expensive and animal-derived components. As cellulose acetate nanofibers is a low-cost cellulose-derived material, that induces cell adhesion and proliferation, we investigated its use associated with a bioactive annatto extract, a food-dye and potential meat preservative, as scaffolds for cultivated meat. Here, the bioactive electrospun nanofibers were evaluated through morphological, mechanical and biological characterizations. The results revealed that the scaffolds were porous with no specific alignment and average fiber diameter of 420±212 nm. Molecular analyzes revealed that in contrast to cellulose acetate scaffold, annatto-loaded cellulose acetate scaffold favor a proliferative state of C2C12 mouse skeletal myoblasts. SEM microscopy images suggests that the nanofiber substrates can sustain long-term culture of the cells, up to 28 days. These results suggest that the combination of cellulose acetate fibers loaded with annatto extract may be an interesting economical alternative for support long-term muscle cells culture with potential application as a scaffold for cultivated meat and muscle tissue engineering.